在长期保存的DNA模板中用双重式PCR检测鼠疫特异性基因的结果分析

Analysis of Detected the Yersinia Pestis DNA in Preserveing Materials for Long-term with the Double Primers PCR Method

目的了解鼠、蚤材料DNA模板长期保存后鼠疫菌特异性基因的检出率。方法应用双重式PCR技术,以鼠疫菌特异性基因Pla和Fra作为引物,进行PCR实验,比较DNA模板在-20℃冰箱保存3d和5年的检测情况。结果实验感染的动物脏器模板,检出阳性率从100%降到25.00%,印鼠客蚤和缓慢细蚤的模板检出阳性率分别从100%降为16.22%和23.33%;现场采集的22份动物脏器模板阳性率从100%降为22.73%;纯菌模板对照阳性率100%。结论鼠疫PCR阳性检出率与模板中的鼠疫菌含菌量及模板保存的时间有关。

Objective This paper reported rhar the double primers （Pla-Fra） PCR ） was detected the Yersinia pesris DNA in preserveing materials of rats and fleas for long -term. Abstract for understanding their positive rare. Methods The double primers（Pla-Fra） PCR was detected the Yersinia pesris DNA in the materials of rats and fleas after preserveing 5 years under -20℃, and with comparing results of 3d. Results The positive rare of rats by rest in- fection was from 100% ro 25.00% and the positive rare of 22 rats from plague prevalent area was from 100% ro 22. 73%. The positive rare of X. cheopis and L. segnis differenenrly were from 100%ro 16.22%and 23.33%. Y. pestis in the meantime was positive. Conclusion The positive rare of the double primers（Pla-Fra-PCR ） was with Y. pesris DNA amount of the materials and preserveing rime.